Cytochemical discrimination between catalases and peroxidases using diaminobenzidine

Roels, Frank; Wisse, Eddie; Prest, B. De; Meulen, J van der

doi:10.1007/bf00490073

Cited by 176 publications

(53 citation statements)

References 120 publications

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“…4). Significant reaction products were only obtained for mitochondrial cristae, most likely caused by an intramitochondrial peroxidase reaction (39). Notably, the DAB-positive organelles adjoined to vacuoles as described for the N. crassa slime mutant were never observed, even when serial ultrathin sections of a cell were inspected (see Fig.…”

Section: Intracellular Distribution Of Catalase Activitymentioning

confidence: 89%

A Eukaryote without Catalase-Containing Microbodies: Neurosporacrassa Exhibits a Unique Cellular Distributionof Its Four Catalases

et al. 2006

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Microbodies usually house catalase to decompose hydrogen peroxide generated within the organelle by the action of various oxidases. Here we have analyzed whether peroxisomes (i.e., catalase-containing microbodies) exist in Neurospora crassa. Three distinct catalase isoforms were identified by native catalase activity gels under various peroxisome-inducing conditions. Subcellular fractionation by density gradient centrifugation revealed that most of the spectrophotometrically measured activity was present in the light upper fractions, with an additional small peak coinciding with the peak fractions of HEX-1, the marker protein for Woronin bodies, a compartment related to the microbody family. However, neither in-gel assays nor monospecific antibodies generated against the three purified catalases detected the enzymes in any dense organellar fraction. Furthermore, staining of an N. crassa wild-type strain with 3,3-diaminobenzidine and H 2 O 2 did not lead to catalasedependent reaction products within microbodies. Nonetheless, N. crassa does possess a gene (cat-4) whose product is most similar to the peroxisomal type of monofunctional catalases. This novel protein indeed exhibited catalase activity, but was not localized to microbodies either. We conclude that N. crassa lacks catalase-containing peroxisomes, a characteristic that is probably restricted to a few filamentous fungi that produce little hydrogen peroxide within microbodies.

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Section: Intracellular Distribution Of Catalase Activitymentioning

confidence: 89%

A Eukaryote without Catalase-Containing Microbodies: Neurosporacrassa Exhibits a Unique Cellular Distributionof Its Four Catalases

et al. 2006

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“…However, this appears to be due to suboptimal preservation of this enzyme under the conditions used. With an adapted methodology (24,29), these bodies become positive. Catalase-positive peroxisomes have already been described by several authors in other yeast species and under other growth conditions (16,22,25,26,28,33,34,39).…”

Section: Discussionmentioning

confidence: 99%

“…In addition, the following variation in the fixation procedure was used for the localization of catalase: the 7.5-,um sections were fixed for 24 h in a 4% formol solution containing 1% CaCl2 (29). After a 1-min wash, the sections were incubated in the catalase medium as described below.…”

mentioning

confidence: 99%

Cytochemical and Biochemical Studies of Yeasts After In Vitro Exposure to Miconazole

Nollin

Belle

Goossens

et al. 1977

Antimicrob Agents Chemother

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Yeast cells exposed to different doses of the antimycotic agent miconazole revealed important cytochemical changes in the topographic distribution of the phosphatases. A strong effect was observed on the behavior of oxidative and peroxidative enzymes. Decreased cytochrome c oxidase and peroxidase activity and increased catalase activity were seen after treatment with a fungistatic dose ofmiconazole, whereas a complete disappearance ofthese enzymes was observed after treatment with a minimal fungicidal dose of miconazole. This was in complete agreement with the quantitative biochemical data. A hypothesis is advanced concerning the possible involvement of peroxidase and catalase in the mechanism of action of this drug., is an imidazole nitrate that has demonstrated broad-spectrum activity against most pathogenic fungi and gram-positive bacteria (7,10,18,19,36). The studies on the uptake and utilization of substances in Candida albicans by Van den Bossche (37) indicated that miconazole induced changes in the permeability of the plasmalemma and the cell wall. At the ultrastructural level, C. albicans cells show changes in a dosedependent manner when grown in the presence of 10`8 to 10-M concentrations of miconazole. These changes range from slight alterations in the cell periphery and increase in the number of peroxisomes and lipid globules, to progressive cytoplasmic deterioration, prominent shape changes, and complete cellular necrosis (11, 12; S. De Nollin, Ph.D thesis, University of Brussels, Brussels, Belgium, 1976). Recent investigations with scanning and transmission electron microscopy (12) on C. albicans cultures treated with a fungicidal dose of miconazole (10-4 M) have shown that the sequence of alterations differs totally from that obtained with lower-dose treatments.The cytochemistry of untreated yeast cells has already been described by several authors (2, 3, 13, 22, 40; S. De Nollin, Ph.D. thesis, 1976). In earlier experiments (16, 13), we tried to optimize the preparatory conditions for the cytochemical examination of yeasts cells by using a modification of the conventional procedure for preservation. This yields, besides a good micromorphology of the cells, sufficient enzyme activities and allows an adequate penetration of substrates and captation ions during the incubation. For the biochemical studies, Van Belle, Goossens, and Van Roy (in preparation) have recently developed a method for obtaining adequate homogenization of the very rigid cells of yeast, thereby yielding reliable enzyme activities that are directly proportional to the number of cells (more than 99.9% of the cells are thoroughly homogenized). Using these modifications of the cytochemical and biochemical preparative techniques, we have tried in this study to evaluate the effects of miconazole on the activities and the distribution pattem of phosphohydrolases and of oxidative and peroxidative enzymes in C. albicans and Saccharomyces cerevisiae. Our particular aim was to see whether there was any correlation between the observed morph...

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“…4, 9), and that in the epithelial cells of the other tissues, such as lacrimal glands and salivary glands (8,20), also produces only a little or no visible cytochemical stain. With respect to the influence of H2O2 on peroxidases, Roels et al (16) have reported the inhibition of peroxidase activity by higher concentration of H2O2. In our experimental conditions, 0.005% H2O2 for the localization of peroxidase yielded the same staining intensity as 0.01% after the technique of The localization pattern of peroxidase-positive cells in the germ-free adult rats was identical to that in the conventional.…”

Section: Discussionmentioning

confidence: 99%

Endogenous peroxidase in the epithelium over peyer's patches of rat small intestine.

Hasegawa

Nishimura

1985

Acta Histochem. Cytochem

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Cytochemical discrimination between catalases and peroxidases using diaminobenzidine

Cited by 176 publications

References 120 publications

A Eukaryote without Catalase-Containing Microbodies: Neurosporacrassa Exhibits a Unique Cellular Distributionof Its Four Catalases

A Eukaryote without Catalase-Containing Microbodies: Neurosporacrassa Exhibits a Unique Cellular Distributionof Its Four Catalases

Cytochemical and Biochemical Studies of Yeasts After In Vitro Exposure to Miconazole

Endogenous peroxidase in the epithelium over peyer's patches of rat small intestine.

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